Betalain compositions and methods therefor

ABSTRACT

Contemplated compositions and methods employ betalains for treatment of various conditions, and especially osteoarthritis, sinusitis, contact dermatitis, acne, an allergic condition, reduced mental alertness, reduced physical strength, reduced physical endurance, and/or impaired mood.

This application is a divisional application of U.S. patent applicationSer. No. 14/726,379, filed May 29, 2015, which is a divisionalapplication of U.S. patent application Ser. No. 13/056,482, filed Jan.28, 2011, issued Jun. 3, 2015 as U.S. Pat. No. 9,060,539, which is anational phase of PCT International Patent Application No.PCT/US09/52293, filed Jul. 30, 2009, and claims priority to our U.S.provisional patent application with the Ser. No. 61/084,879, filed Jul.30, 2008, and to our U.S. provisional patent application with the Ser.No. 61/140,541, filed Dec. 23, 2008, both of which are incorporated byreference herein.

FIELD OF THE INVENTION

The field of the invention is compositions and methods for treatment ofconditions in human, and especially to as they relate tobetalain-containing compositions for treatment of osteoarthritis, acne,allergic conditions, sinusitis, and contact dermatitis. Compositions andmethods contemplated herein also relate to use of thebetalain-containing compositions as a non-caffeine stimulant.

BACKGROUND OF THE INVENTION

Osteoarthritis (OA) is a very common condition, and it is expected thatabout 80% of the U.S. population will have radiographic evidence of OAby age 65. Osteoarthritis is characterized by loss of articularcartilage that is frequently accompanied by pain and swelling of thetissue proximal to the affected joint, which in turn often leads tolocal or regional atrophy of muscles associated with that joint. WhereOA has no identified underlying cause, OA is also referred to as primaryor degenerative OA, while secondary OA is typically precipitated byvarious disorders or diseases (e.g., diabetes, local injury orinfection, joint instability, etc.). In contrast, rheumatoid arthritis(RA) is a chronic, systemic autoimmune disorder in which the immunesystem attacks the affected joint, leading to significant inflammationand subsequent tissue degeneration.

Due to their different etiologies, RA and OA have distinct therapeuticapproaches. For example, RA is typically treated with cyclosporine,methotrexate, or penicillamine, and is more severe cases with TNF-alphaor IL-1 blockers. To reduce inflammation, glucocorticoids and/ornon-steroidal anti-inflammatory can be administered. On the other hand,OA is often treated with physical therapy, weight management, and/orvarious nutritional supplements. Among other supplements,methylsulfonylmethane, glucosamine and/or chondroitin sulfate havegained significant attention as supplements. More recently, variousplant-derived supplements were described for treatment of OA. Forexample, WO 05/053710 teaches use of a drying process in the manufactureof glucosamine from various plant materials. In another example, a plantextract for treatment of osteoarthritis is prepared from Morindacitrifolia as described in U.S. Pat. App. 2007/0196527.

Most currently available energy drinks (e.g., Red Bull™, Rockstar™, JavaMonster™, etc.) typically include significant quantities of caffeine andother methylxanthines, selected B vitamins (e.g., B3, B6, B12) and smallmolecule effectors (e.g., taurine, creatine, maltodextrin,glucuronolactone, etc.), as well as various herbal extracts/preparations(e.g., guarana, ginseng, ginkgo, yerba mate, etc.). Additionally, atleast some of the currently marketed energy drinks also includesubstantial quantities of sugar.

Most of the stimulatory effect of such energy drinks is derived fromcaffeine, which is present in the average 8 ounce serving in an amountof about 80 mg. While an 80 mg dose of caffeine is generally notproblematic for the average consumer, multiple servings of energy drinkstend to produce undesirable, and in some cases even dangerous sideeffects. For example, caffeine dosages above 250 mg tend to producenervousness, irritability, sleeplessness, increased urination,arrhythmia, gastric irritation, and even loss of bone mass.Unfortunately, elimination of caffeine from the system typically resultsin a ‘bummer’ or crash, and sudden drops in caffeine levels from largedoses have even been associated with seizures.

To overcome at least some of the problems associated with ingestion oflarge quantities of caffeine, other energy drinks are currently marketed(e.g., 5-Hour Energy®) that include large quantities of B-vitamins,amino acids (tyrosine, phenylalanine), taurine, glucuronolactone andonly small quantities of caffeine. While such energy drinks typicallyavoid subjective discomfort upon caffeine elimination, other sideeffects are often encountered, including niacin flush and sleeplessness.Moreover, long-term effects of high-dose vitamin B administration isgenerally unknown.

Red beets have long been a common source of various nutrients, andparticularly of sugar and betaine (i.e., trimethylglycine). Furtheruseful compounds obtained from red beet include betalains, whichrepresent a chemically diverse group of red to violet coloredbetacyanins (e.g., amaranthin, isoamaranthin, etc.) and typicallyyellow-colored betaxanthins (e.g., vulgaxanthin), which have found useas pharmaceutical and food coloring agents and as antioxidants. Forexample, betalains have been reported as antioxidants (see e.g., J AgricFood Chem. 2001 November; 49(11): 5178-85), which may have particularrelevance to prevention of LDL cholesterol oxidation (see e.g., FreeRadic Res. 2003 June; 37(6):689-96). More recently, as described in ourcopending International application WO 08/094705, betalain compositionswere also found to be effective in reducing serum triglycerideconcentration, inhibiting NF-kB, and stimulation of SIRT. Furthermore,red beet fiber has been reported to have numerous desirable propertieswhen ingested, including reduction of total cholesterol and triglyceridelevels, while increasing HDL cholesterol.

However, despite the numerous advantages of red beets and red beetextracts, red beets have not been reported as containing activecompounds for treatment of OA, various allergic conditions or conditionsassociated with allergies, and lack of energy. Complicating matters withred beet extracts is their problematic manufacture and dispensation,especially where the red beet preparation is in dry form. Among otherdifficulties, betalains prepared from freeze-dried beet juice are oftenclumpy and highly hygroscopic. Therefore, such preparations arenotoriously difficult to weigh out and aliquot, especially whererelatively small quantities are distributed. Worse yet, most of thecurrently known dry betalain preparations are limited to betalainconcentrations of about 1 wt % (total betalains), and almost all of theattempts to increase the betalain concentration by extraction or othermeans also leads to an increase of hygroscopicity and clumping.Alternatively, where red beet extracts are in liquid form, such extractsare often highly instable and tend to degrade rapidly.

Therefore, while numerous compositions and methods of red beetpreparations are known in the art, all or almost all of them suffer fromdisadvantages. Consequently, there is still a need to provide improvedcompositions and methods for red beet preparations.

SUMMARY OF THE INVENTION

The present invention is directed to beet-derived compositions andmethods in which such compositions are used for treatment of variousconditions, and particularly osteoarthritis, acne, contact dermatitis,sinusitis, and/or conditions associated with allergy. Most preferably,contemplated compositions and methods will be enriched in betalains andhave a reduced sugar content as compared to unprocessed beet juice.

In one preferred aspect of the inventive subject matter, a method ofproviding a product for human consumption will include a step ofproviding or including a quantity of betalains in a product for humanconsumption, wherein the quantity is effective to reduce at least onesymptom associated with at least one of osteoarthritis, sinusitis,contact dermatitis, acne, an allergic condition, reduced mentalalertness, reduced physical strength, reduced physical endurance, andimpaired mood when the product is ingested at a recommended dosage andschedule. In a further step, a person is then instructed to orallyadminister the product at the recommended dosage and schedule to soreduce the at least one symptom associated with the condition. Mosttypically, the instruction will further inform the person that it is thequantity of betalains that is effective in the reduction of thesymptom(s).

While not limiting to the inventive subject matter, it is generallypreferred that the quantity of betalains is prepared from red beet,and/or that the quantity of betalains is included into the product as asolid preparation comprising total betalains at a concentration of atleast 2 wt %, and more typically at least 10 wt %. It is also generallypreferred that the solid preparation has a betalain to sugar ratio of atleast 0.3, and more typically of at least 1.0. It is still furthergenerally preferred that the dosage and schedule are selected such thatthe total daily intake of the composition provides between 10 mg and 50mg total betalain. Thus, in especially contemplated aspects, the productis formulated as a nutritional supplement, wherein the supplement isformulated to provide between 10 and 250 mg of total betalains in adaily dose in which the supplement has a weight of less than 1000 mg.

Consequently, in another preferred aspect of the inventive subjectmatter, a method of treating osteoarthritis will include a step ofadministering to a person in need thereof betalains at a dosage andschedule effective to reduce at least one symptom associated withosteoarthritis. Most preferably, the dosage and schedule are selectedsuch that the total daily intake of the betalains is between 10 mg and50 mg, and where desired, the betalains are present in a compositionisolated from a plant material wherein the composition has a betalain tosugar ratio of at least 1.0.

In a still further preferred aspect of the inventive subject matter, amethod of providing a nutraceutical stimulant will have a step ofproviding betalains in the nutraceutical stimulant in a quantityeffective to increase at least one of mental alertness, physicalstrength, physical endurance, and mood when the nutraceutical stimulantis ingested at a recommended dosage and schedule, and in another step, aperson is instructed to orally administer the stimulant at therecommended dosage and schedule to so increase at least one of mentalalertness, physical strength, physical endurance, and mood. It isgenerally preferred that the dosage and schedule are selected such thatthe total daily intake of the stimulant provides between 10 mg and 50 mgbetalains, and/or that the betalains are added to the stimulant in acomposition that is isolated from a plant material and that has abetalain to sugar ratio of at least 1.0.

Therefore, and viewed from a different perspective, use of abetalain-containing preparation is contemplated in the manufacture of anorally administered composition for treatment of osteoarthritis,sinusitis, contact dermatitis, acne, an allergic condition, reducedmental alertness, reduced physical strength, reduced physical endurance,and/or impaired mood. In especially preferred aspects, thebetalain-containing preparation is a dried preparation having a betalainconcentration of at least 5 wt %, and/or the betalain is included in thepreparation in an amount of at least 15 mg per dosage unit.

Various objects, features, aspects and advantages of the presentinvention will become more apparent from the following detaileddescription of preferred embodiments of the invention.

DETAILED DESCRIPTION

The inventors have discovered that betalain-containing preparations, andespecially various red beet preparations are suitable to reduce one ormore symptoms associated with certain conditions, and especiallyosteoarthritis, sinusitis, contact dermatitis, acne, an allergiccondition, asthma, reduced mental alertness, reduced physical strength,reduced physical endurance, and/or impaired mood.

Most preferably, the compositions and methods according to the inventivesubject matter are orally administered such that a daily dosage of totalbetalains of between 10 mg and 250 mg is achieved. While preferredadministration of the compositions contemplated herein is not restrictedto a specific schedule or dosage, it is generally preferred that thecompositions are administered over a period of at least 3 days, and moretypically of at least 2 weeks, and most typically at least 1 month usingoral administration preferably between once daily and three times daily.

In one exemplary preferred aspect it is contemplated that the beetpreparation is prepared from red beet juice in an extraction processthat produces a dry material at a concentration of betalains of at least2 wt %, and more typically at least 5-10 wt % (e.g., see sectionexamples and experiments below). Particularly preferred compositions aredescribed and known from our International patent application WO08/094705 (serial number PCT/US08/01418), which is incorporated byreference herein. Such preparation is then filled in capsules for oraladministration of about 100 mg total preparation per capsule and orallyadministered three times daily over a period of at least three weeks.

For example, a betalain-containing composition was prepared following aprotocol substantially as described in WO 2008/094705, which isincorporated by reference herein. Briefly, commercially available beetjuice (about 65 brix; obtained from SVZ International) having a totalbetalain content of about 0.6 wt % on dry basis was filtered to removeparticulates and the filtrate was used without further modification forchromatography. The filtrate was passed through a column packed with ahydrophobically modified styrene resin (commercially available as ResinHST-226 from VDF Futureceuticals) at between 1-50 bed volumes per hourat a loading with between 1-20 bed volumes. Pass fractions werediscarded, and betalains were eluted from the resin using a mild bufferat slightly basic pH (e.g., 0.1-0.2 M ammonium acetate in water, pH 8.2to 8.4, at a temperature of 110° F., or 0.1-0.2 M ammonium carbonate inwater at pH range 7.0 to 9.0, same temperature). The so obtained eluatewas freeze-dried without further modification to a dry product. Samplesprepared according to the protocol above had a typical content asprovided in Table 1 below:

TABLE 1 Compound Quantity Total betalains 27 wt %  Betaine 30 wt % Amino acids 4 wt % Fiber 2 wt % Betaine 30 wt %  Water 8 wt %Undetermined 23 wt % 

It should be noted that the so prepared composition had a relativelyhigh nitrogen content as measured by Kjeldahl and Folin-Ciocalteu.Notably, total nitrogen was present in the form of betaine(trimethylglycine), amino acids (which are most likely a mixture of freeamino acids and amino acids covalently bound to one or more betalains),free ammonium ions and ammonium ion pairs as well as ammoniatedcompounds (and especially aminated betalains). Due to the relativelymild and rapid isolation process, it is also contemplated that the soisolated betalains have a relatively large fraction of carboxylatedbetalains (e.g., at least 50 mol %, more typically at least 80 mol %,most typically at least 90 mol % of total betalains, with17-decarboxy-forms less than 10 mol %, more typically less than 5 mol %,and most typically less than 3 mol % of total betalains), all or atleast some of which may contribute to the observed activities.

In still further contemplated aspects, it should be appreciated thatnumerous alternative preparations of betalain-containing compositionsare deemed suitable and include those in which the total betalainconcentration is at least 2 wt %, more preferably at least 5 wt %, evenmore preferably at least 10 wt %, and most preferably at least 20 wt %,typically at a betalain to sugar ratio of at least 0.3, more preferablyat least 1.0, and most preferably at least 2.0. Moreover, it isgenerally preferred that the betalain-containing compositions willreadily and substantially completely dissolve in water at fairly highconcentrations (typically at least 90%-98% at 50-100 mg/ml). Stillfurther it should be appreciated that while it is generally preferredthat the betalain-containing compositions will be dried preparations,gels, and liquid preparations are also deemed suitable for use herein.Such preparations may be further processed to achieve a particular pH,consistency, or concentration of non-betalain components. In yet furtheraspects of the inventive subject matter, the betalains in thebetalain-containing composition have a near natural composition (i.e.,no individual betalain will be under- or over-represented by at least10% as compared to composition prior to isolation).

Therefore, numerous other manners of producing betalain compositions arealso deemed suitable and may use various materials as startingmaterials, including comminuted beet root or root peelings, root cellsuspension culture, cactus pears, and even selected higher fungi. Forexample, where relatively low yields are acceptable, WO 98/26792describes various methods of preparing betalain extracts from root pulp,while lyophilized beet is used as starting material as described in US2003/0036565, which is subsequently ground, solvent extracted, andsubjected to crosslinked dextran chromatography to yield distinctbetalain fractions. Alternatively, beet juice concentrate may be spraydried to a powder comprising about 0.4 wt % betalains.

In other known methods (e.g., as described in U.S. Pat. No. 4,238,518 orGB patent 1 559 275), stabilized betanidine extracts are prepared usingion exchange chromatography while Garin et al. describe in U.S. Pat. No.4,409,254 a process in which beet root extract is subjected at very lowpH to chromatography using a non-ionic resin to so produce aconcentrated eluate. Further suitable processes and products aredescribed in U.S. Pat. No. 4,027,042 in which beet juice or beet pulp issubjected to a yeast fermentation and subsequent work-up.

Alternatively, it should be noted that numerous raw or startingmaterials other than red beet juice are also deemed suitable, andespecially contemplated materials include raw red beet root or portionsthereof (e.g., in solid, macerated, or paste form), red beet processingwaste liquids, and red beet root cell cultures and culture supernatants.In still further contemplated aspects, suitable starting materials mayalso include plant materials that comprise betalains as a natural (orrecombinantly produced) pigment. Therefore, betalain containing plantswill also include those found in the order of the caryophyllales andselected basidiomycota, and in various cacti (e.g., prickly pear cactusand related plants). Consequently, it should be appreciated thatdepending on the type of starting material the exact composition ofbetalains may vary considerably. Additionally, it should be appreciatedthat the compositions according to the inventive subject matter may alsocomprise one or more individual isolated or synthetic betalains as soleor supplemental ingredient. However, it is generally preferred that thebetalains in the preparation are a complex mixture of betalains withnatural or near natural (i.e., deviation of each betalain of less than10% as compared to natural composition) relative proportions.

In typical examples according to the inventive subject matter, thebetalain-containing preparation has a total betalain concentration of atleast 4.0 wt %, more typically at least 10.0 wt %, even more typicallyat least 15.0 wt %, and most typically at least 20.0 wt %. It should benoted that the chemical composition of the betalain preparationsaccording to the inventive subject matter is a complex composition thatincludes a plurality of chemically distinct betalains. Thus, the complexpreparation will include both betaxanthins and betacyanins. Mostpreferably, the betalain preparations presented herein have a nearnatural composition (i.e., no single betacyanin or betaxanthinoriginally present in the red beet is concentrated or depleted more than30% [and more typically more than 15-20%] relative to the naturalcomposition). Analysis of betalains and complex compositions can beperformed as described by Corke et al (J Chromatogr Sci. 2005 October;43(9):454-60) or Pourrat et al. (Journal of Food Science 53 (1),294-295). Thus, typical preparations will include at least ten, moretypically at least twenty, and most typically at least 25 chemicallydistinct betalains. Therefore, it should be noted that for various usesmost commercially available betalain-containing preparations (e.g.,juices, juice concentrates, powders) are also deemed suitable, althoughsignificantly higher overall quantities of those preparations may beneeded (which in many cases will increase the dietary intake of sugars).

Depending on the particular source material and solvents used, it shouldbe noted that the betalain to sugar ratio in contemplated compositionsis least 0.3, more typically at least 0.5, even more typically at least1.0, and most typically at least 2.0. Viewed from a differentperspective, most preferred preparations will have a betalain to sugarratio between 1.0 and 5.0, and even more typically between 2.5 and 4.5.With respect to the remaining sugars in contemplated compositions itshould be recognized that the chemical nature will vary and depend onthe starting material and work-up. However, most typical remainingsugars include may be mono-, oligo, and/or polysaccharides, sugaralcohols, and pectins. Thus, it should be appreciated that where theoligo, and/or polysaccharide concentration is to be reduced, enzymaticor fermentative processes may be used to achieve such reduction.Alternatively, or additionally, residual monosaccharides may be removedusing various manners known in the art, including ultrafiltration,molecular sieving, and/or enzymatic conversion.

With respect to the quantity of the beet preparation administered in adaily dose it is generally preferred that the amount of the preparationis between 1 mg and several hundred grams (or even more). However, andwhile not limiting the inventive subject matter, it should be noted thatthe preparations will typically be administered in an amount that isneeded to provide an effective daily dose of betalains to reduce atleast one symptom of the condition. Typically, the effective dose willbe in the range of about 1 mg to about 1000 mg, more typically in therange of about 10 mg to about 500 mg, and most typically in the rangeabout 25 mg to about 250 mg.

It is further contemplated that the preparations according to theinventive subject matter will be solid betalain-containing preparationsthat may be formulated in numerous manners. For example, suitableformulations include oral formulations (e.g., tablets, capsules,dragees, ready-to-mix formulation, etc.) in which the preparation is theprimary ingredient or formulations in which the preparation is disposedin an edible carrier (e.g., bar, snack, confectionery item, etc.).Alternatively, the betalain-containing preparation may also be a liquidformulation in which the liquid is mixed or encapsulated in anutritionally acceptable carrier. For example, suitable liquidformulations may include drinks, syrups, gelatin-encapsulated liquidextracts, oil infusions, soft gels, coatings for capsules, and liquidtinctures. Moreover, and especially where only a single betalain is usedas the active ingredient, it is contemplated that the betalain may beformulated in a pharmaceutically acceptable formulation for oral orparenteral administration. In such case, especially suitableformulations include oral formulations, topical formulations, andformulations for injection.

Additionally, it should be appreciated that contemplatedformulations/compositions for administration may also include a secondcompound that is known or reported to reduce symptoms of OA or symptomsof other conditions. For example, especially suitable second compoundsinclude glucosamine, methylsulfonylmethane, and chondroitin sulfatewhere the preparation is provided as a nutritional supplement. On theother hand, where the preparation is provided as a pharmaceuticalcomposition, the second compound may be a non-steroidalanti-inflammatory drug, a COX-inhibitor, etc. Consequently, contemplatedcompositions especially include those in which a single betalain or aplurality of chemically distinct betalains are formulated as an orallyadministrable composition, optionally in combination with anothercompound that is known or reported to reduce symptoms of OA. Suchcompositions may be formulated, for example, as a tablet or capsule,optionally in further combination with one or more vitamins, minerals,or herbal preparation, or as an edible product that is fortified withthe betalain(s). For example, such products may be a snack, a bar, acandy, a drink, etc.

Therefore, administration of contemplated compositions is preferablyoral using a daily dosage that contains about 1-250 mg, and mostpreferably between 10 and 100 mg of one or more betalains. Theimprovements in energy and alertness are observed in most cases after 3days of continuous administration, whereas improvements inosteoarthritis and other conditions is typically observed after about 1week. In most cases, administration will typically be between once dailyand four times daily (or even more) with an amount of typically between10 and 250 mg per dose. Therefore, especially preferred dosages will bebetween about 0.01 mg/kg to about 10 mg/kg (most preferably 0.1 mg/kg toabout 1 mg/kg) body weight. While long-term administration is generallypreferred, it should be appreciated that administration may extend overat least 3 days, more typically at least 1 week, even more typically atleast 1 month, and most typically between 1 month and 6 months.Interestingly, discontinued use of contemplated compositions was notassociated with any withdrawal symptoms normally found with energydrinks.

Experiments and Examples Exemplary Processes for Extraction

Option 1: Extraction started with commercially available beet juice(about 65 brix; obtained from SVZ International) having a total betalaincontent of about 0.6 wt % on dry basis. The juice was filtered to removeparticulates and the filtrate was used without further modification forchromatography. A column was packed with a hydrophobically modifiedsilica resin (commercially available as Resin HSI-564 from VDFFutureceuticals; Momence, Ill. 60954), and the filtered juice was passedthrough the column at between 1-50 bed volumes per hour at a loadingwith between 1-20 bed volumes. The pass fractions were discarded, andthe betalains were eluted from the resin using a mild buffer at slightlybasic pH (e.g., 0.1-0.2 M ammonium acetate in water, pH 8.2 to 8.4, at atemperature of 110° F., or 0.1-0.2 M ammonium carbonate in water at pHrange 7.0 to 9.0, same temperature). The so obtained eluate wasfreeze-dried without further modification to a dry product. Quantitativeanalysis of the product revealed a total betalain content of about 15 wt% at 6 wt % total sugar and a residual water content of about 8 wt %.The dry product was then ground using a rotating blade grinder to form apowder product that was passed through a 60 mesh sieve. More than 90%,and more typically more than 95% of the free-flowing dry productdissolved within less than 2 minutes at a concentration of between 0.1mg/ml and 15 mg/ml, more typically at a concentration of between 10mg/ml and 50 mg/ml, and more typically at a concentration of between 50mg/ml and 100 mg/ml (and in some cases even higher, such as between 100mg/ml and 200 ml/ml). Even more remarkably, the product was stable overat least 2 weeks storage at 75° F. and 50% relative humidity without anyobservable changes in composition, free-flowability, or otherparameters, and did not aggregate to larger particles or clumps. Indeed,after 2 weeks (and longer) storage at the defined condition,substantially the same amount (>90%) of the dry powder passed through amesh sieve of same mesh size.

Option 2: Extraction started as above with commercially available beetjuice (about 65 brix; obtained from SVZ International) having a totalbetalain content of about 0.6 wt % on dry basis. The juice was filteredto remove particulates and the filtrate was used without furthermodification for chromatography. A column was packed with ahydrophobically modified styrene resin (commercially available as ResinHST-226 from VDF Futureceuticals) and the filtered juice was passedthrough the column at between 1-50 bed volumes per hour at a loadingwith between 1-20 bed volumes. The pass fractions were discarded, andthe betalains were eluted from the resin using a mild buffer at slightlybasic pH (e.g., 0.1-0.2 M ammonium acetate in water, pH 8.2 to 8.4, at atemperature of 110° F., or 0.1-0.2 M ammonium carbonate in water at pHrange 7.0 to 9.0, same temperature). Without drying as described above,the so obtained eluate was mixed with a water-soluble polysaccharide(e.g., to about 30 wt % maltodextrin final concentration) as carrier andspray-dried to a dry product having about 10 wt % betalains at 45 wt %total sugar content with a residual water content of about 5 wt %. Theaverage particle size of the spray-dried product was about 150 micron(passed through 60 mesh sieve). More than 90%, and more typically morethan 95% of the free-flowing dry product dissolved within less than 2minutes at a concentration of between 0.1 mg/ml and 15 mg/ml, moretypically at a concentration of between 10 mg/ml and 50 mg/ml, and moretypically at a concentration of between 50 mg/ml and 100 mg/ml (and insome cases even higher, such as between 100 mg/ml and 200 ml/ml). Asabove, the product was stable over at least 2 weeks storage at 75° F.and 50% relative humidity without any observable changes in composition,free-flowability, or other parameters, and did not aggregate to largerparticles or clumps. Once more, after 2 weeks (and longer) storage atthe defined condition, substantially the same amount (>90%) of the drypowder passed through a mesh sieve of same mesh size.

Analytic Tests

Betalain-rich red beet extract, (RBE) was prepared following the methodsdescribed in Options 1 and 2 above.

Proximate Analysis. Moisture content of RBE was determined according toUSP loss-on-drying (LOD) method. Sample was heated in a vacuum oven at70° C. for 7 hours. Total protein content was determined based onBradford method. Available carbohydrates were calculated by deductingthe sum of crude protein, crude fat, ash and moisture from 100% of theDM. Ash content was determined by igniting the sample at 550° C. inelectric furnace, AOAC 923.03 (AOAC, 2005).

Mineral Analysis. The 1.2 g sample test portion was dry ashed at 500°C.±50° C. for 8 hours and treated with HNO₃. The resultant ash wastreated with concentrated hydrochloric acid (5%), dried, and redissolvedin hydrochloric acid solution (16). The amount of each element (Al, Ba,B, Ca, Cu, Fe, K, Mg, Mn, Na, Zn) was determined by comparing theemission of the unknown sample against the emission of each element fromstandard solutions using Inductively Coupled Plasma Atomic EmissionSpectroscopy (ICAP-61E-Trace, Thermo Jarrell-Ash). All standardsolutions used were obtained from Inorganic Ventures (Christiansburg,Va.—USA) and were of analytical-reagent grade.

Sugar Analysis. The sugars (sucrose, glucose, fructose, maltose,lactose, and galactose) were extracted from an accurately weighed samplewith 80% ethanol by allowing it to stand for 24 hours with occasionalswirling. Aliquots were dried under inert gas and reconstituted with ahydroxylamine hydrochloride solution in pyridine containingphenyl-β-D-glucoside as the internal standard. The resulting oximes wereconverted to silyl derivatives with hexamethyldisilazane (HMDS) andtrifluoracetic acid (TFA) treatment and subsequently analyzed by gaschromatography using a flame ionization detector.

Total Dietary Fiber Analysis. Determination of total dietary fiber (TDF)was based on the methods of. Duplicate samples were cooked at ˜100° C.with heat stable α-amylase to give gelatinization, and then digestedwith enzymes in a phosphate buffer to break down starch and someprotein. Ethanol was added to each sample to precipitate any solublefiber. The samples were filtered, and residues were rinsed with ethanoland acetone to remove starch and protein degradation products andmoisture. Protein content was determined for one of the duplicates; ashcontent was determined for the other. The total dietary fiber in thesample was calculated after adjustment for the protein and ash values.

Amino Acid Analysis. A sample of RBE was hydrolyzed in hydrochloric acid(HCl) and adjusted to pH 2.2 for all amino acids except tryptophan.Tryptophan samples were hydrolyzed in sodium hydroxide and adjusted topH 5.2. Individual amino acids were determined by comparison using anautomated amino acid analyzer.

Fatty Acid Profile Analysis. Fat and fatty acids were extracted from RBEsample by hydrolytic method based on (23). Pyrogallic acid was added tominimize oxidative degradation of fatty acids during analysis. Atriglyceride, triundecanoin (C11:0), was added as internal standard. Fatwas extracted into ether, then methylated to fatty acid methyl esters(FAMEs) using BF3 in methanol. FAMEs were quantitatively measured bycapillary gas chromatography (GC) against a C11:0 internal standard.Total fat was calculated as sum of individual fatty acids expressed astriglyceride equivalents. Saturated and monounsaturated fats werecalculated as sum of respective fatty acids.

Vitamin Analysis. Vitamin C in the RBE sample was extracted, oxidized,and reacted with o-phenylenediamine to produce a fluorophor. The vitaminC content was determined by comparison of the sample extractfluorescence to the fluorescence of known standard. Beta caroteneanalysis was performed by HPLC method.

Total Betalain Quantification. Quantification of betalains was performedby a spectrophotometric multiple-component method of Nilsson using aUV-VIS spectrophotometer Shimadzu 1650 PC (Shimadzu Corporation, Kyoto,Japan). The determination of pigment concentration (i.e., betacyaninsand betaxanthins) was calculated in terms of betanin and vulgaxanthin-I,respectively. The total pigment content (betalain) was expressed as thesum of betacyanin and betaxanthin components. Pigment contentcalculations were based upon the absorptivity values A^(1%) 1120 forbetanin and 750 for vulgaxanthin-I.

Betalain Profile Analysis. 20 mg of RBE was shaken with 1 ml of waterfor 10 min under neutral gas in a 2 ml glass vial. Samples werecentrifuged and supernatant was analysed directly by LC-DAD or LC-MSwithout any purification. A Gynkotek HPLC system with UVD340U, GynkotekHPLC pump Series P580 and thermostat (Gynkotek Separations, H. I.Ambacht, The Netherlands) was used for chromatographic analysis. Theanalytical column was a Luna C-18(2) 250×3 mm I.D., 5 μm (Phenomenex,Torrance, Calif., USA). The following gradient system was used for theseparation of analytes: 3% A in B at 0 min; gradient to 16% A in B at 17min; gradient to 50% A in B at 30 min (A, acetonitrile; B, 2% formicacid in water). The injection volume was 10 μL and the flow rate of 0.5mL/min was applied. Detection was generally performed at λ=538 nm with aUV-Vis detector or a DAD (diode array detection) system at 533, 505, and480 nm, respectively (27). The columns were thermostated at 35° C.Positive ion electrospray mass spectra were recorded on a ThermoFinniganLCQ Advantage mass spectrometer (San Jose, Calif., USA) at electrosprayvoltage 4.5 kV, capillary 250° C. and sheath gas: N₂ coupled to aThermoFinnigan LC Surveyor pump applying in HPLC gradient System 1. TheMS was controlled, and total ion chromatograms and mass spectra wererecorded using ThermoFinnigan Xcalibur software (San Jose, Calif., USA).

Clinical Study Description and Design

This study was designed to be an open type clinical discovery ratherthan a clinical efficacy study. The primary goal was to verify whetherRBE may improve pain and fatigue associated with osteoarthritisconditions. The secondary goal of this study was to identify MinimumEffective Dose (MED). Therefore, the inventor employed a multiplefixed-dose type study with three time points: (Day 1, 5, 10). There were8 subjects per experimental group. Each group was treated with 100 mg(Group 1), 70 mg (Group 2) or 35 mg (Group 3) twice per day. Allparticipants were asked to take one capsule of RBE 30 min prior toeating a meal.

Subjects for this study were selected randomly from a group of peoplewho had been previously diagnosed with Osteoarthritis and who hadreported symptoms characteristic of OA such as joint pain, limited jointflexibility, and feeling energy-depleted due to chronic pain and jointproblems. The inventor used McGill pain score system and Energy Scoresystem questionnaires at day 1, 5 and 10 as a means of quantifyingsymptoms.

Recruitment of subjects, treatments, blood sampling at day 1, 5 and 10,Mc-Gill and Energy score tests and blood chemistry were performed byNutra Clinical, Inc. (San Diego, Calif., USA). Measurements of humancytokines and chemokines in collected sera were provided by Quensys,Inc. on research service basis. AOPP testing was performed on seracollected from subjects at day 1 and 10 days after the treatment inFutureCeutical's lab using commercially available kit (Cell Biolabs,Inc, USA).

Treatment of volunteers with contemplated compounds as used for OA forcontact dermatitis, eczema, acne, and sinusitis was performed by twicedaily administration of 100 mg of the above composition with subjectiveevaluation s indicated below.

Results

Nutritional composition of RBE. Red beet (Beta vulgaris L.) is commonlyconsumed as food product and FutureCeuticals' RBE is a speciallyprocessed extract obtained from this material. In Table 2 below, RBE isbriefly compared with four different red beet products manufactured byother known concentration techniques. Due to the inventor's productionprocess, the RBE material is depleted of sugars and enriched in totalbetalains up to 24%. More detailed basic chemical information about RBEis presented in Table 3.

TABLE 2 Concentration [%] Glu- Mal- Su- Total Total Sample cose Fructosetose crose sugars betalains RBE <0.1 <0.1 <0.1 0.2 0.5 24.6 Freeze DriedBeet 1.15 0.91 1.08 48.5 51.7 3.06 Spray Dried Beet 1.93 0.84 ND^(a)29.8 32.6 0.43 Air Dried Beet 0.71 0.74 ND 42.7 44.1 0.59 Beet Juice1.62 1.42 ND 41.3 44.4 1.65 Concentrate

TABLE 3 Concentration [mg/100 g] Proximal constituents Moisture 8.8Protein 6.5 Total Fat 0.014 Monosaturated Fatty Acids 0.012 Trans FattyAcids <0.007 Polyunsaturated Fatty Acids <0.007 Saturated Fatty Acids<0.007 Ash 2.04 Total Carbohydrates 13.8 Dietary Fiber 3.57 Protein byBradford method 6.51 Calories [cal/100 g] 375 Vitamins Vitamin A [IU/100g] <35.0 Beta Carotene [mg/100 g] <0.02 Vitamin C [mg/100 g] <1.0 Aminoacids Aspartic Acid 132 Cystine 195 Tryptophan 644 Methionine 67.9Threonine 49.2 Serine 57.2 Glutamic Acid 741 Proline <10.0 Glycine 297Alanine 59.1 Valine 57.1 Isoleucine 26.3 Leucine 39.8 Tyrosine <10.0Phenylalanine 44.7 Lysine 61.8 Histidine 45.6 Arginine 386 Mineralelements Al 3.71 Ba 0.73 B 1.21 Cu 3.28 Fe 28.9 Mg 152.0 Mn 13.4 K 50.6Ca 74.8 Na 43.5 Zn 13.3

Betalain Composition.

The RBE sample was chromatographically analyzed to obtain a basicbetalain profile (Table 4; ^(a)Peak area per injected sample measured atλ_(max). Orientative data for relative comparison between the pigmentsignals). Because the profiles of the compounds can change during apurification process, and due to the possibility that some pigmentdegradation might occur, it was decided to analyze the samples directlyafter extraction with water. Only highly abundant basic compounds werelisted in Table 4.

TABLE 4 Peak area per injected sample Peak Retention measured at λ_(max)^(a) m/z No. Compound time [min] (×10⁻⁵) λ_(max) [M + H]⁺ 1 Betanin 16.912.6 535 551 2 17-decarboxy- 17.5 0.60 505 507 betanin 1′ Isobetanin18.2 15.6 535 551 2′ 17-decarboxy- 19.2 0.44 506 507 isobetanin 3Neobetanin 22.3 3.9 464 549

Besides the highest peaks of betanin/isobetanin 1/1′, a prominent peakof neobetanin 3, which had been frequently detected in Beta vulgaris L.roots, was also observed in the chromatogram. The presence of excessiveamounts of 3 in betacyanin-bearing samples had been attributed manytimes to the degradation of betanin during processing of the samples;however, in this study, the pre-concentration factor was similar to thatof betanin/isobetanin (data not shown) suggesting that the presence of 3was rather a result of its enrichment than dehydrogenation of betanin.This effect is under investigation. Interestingly, the concentration ofboth diastereomers of betanin/isobetanin 1/1′ was similar, indicating apossible epimerization during the whole process of extractconcentration. The principal pigments 1/1′ were mostly responsible forthe total betalain content (24.6%) measured spectrophotometrically.

Further inspection of the chromatograms and mass ion traces revealedalready known 17-decarboxylated betanins 2/2′ present at lower levels inthe concentrated extract. In comparison to the starting material, theirenrichment factor was much higher than in the case of betanin (data notshown), which obviously could be a result of betanin degradationprocess. The presence of 2/2′ has been frequently attributed todecarboxylation during heating of betanin solutions (27, 31-33);however, the degradation at room temperature cannot be excluded.

Effect of RBE on Pain Feelings as Measured Using McGill Test.

This study was performed to verify our hypothesis that betalain-richfood-grade material RBE may reduce discomforts associated with painfuland swollen joints in people suffering Osteoarthitis. Collected resultsshow that all subjects reported reduced pain level in the dose-dependentmanner as measured by using McGill Questionnaire. Detailed data areprovided in Table 5. Following these data, it is clearly noticeable thattreatment with RBE resulted in a significant improvement of the Sensorypart of McGill.

TABLE 5 McGill Scores Group Sensory Part Affective Part Evaluative PartDose Day 1 Day 5 Day 10 Day 1 Day 5 Day 10 Day 1 Day 5 Day 10 Group 1 -100 mg Average 58.87 41.00* 40.00* 4.12 3.62 2.25* 16.37 15.62 15.87 StD3.31 4.40 7.21 1.80 2.19 1.48 3.42 4.83 4.45 Max 63 47 51 6 6 4 20.0021.00 21.00 Min 53 35 30 1 1 1 10.00 8.00 8.00 StE 1.17 1.55 2.54 0.630.77 0.52 1.20 1.71 1.57 Sum^(a) 471 328 320 33 29 18 131 125 127 McGillDay 1: 79.36; Day 5: 60.24; Day 10: 58.12 Score Group 2 - 70 mg Average59.62 44.37* 35.5* 2.25 2.00 1.75 15.50 13.37 14.75 StD 5.57 10.68 4.722.31 1.69 0.70 3.89 3.96 2.81 Max 68 63 46 6 6 3 23 19 18 Min 53 35 32 11 1 12 9 10 StE 1.97 3.77 1.66 0.81 0.59 0.25 1.37 1.40 0.99 Sum 477 355284 18 16 14 124 107 118 McGill Day 1: 77.37; Day 5: 59.74 Day 10: 52.00Score Group 3 - 35 mg Average 63.25 56.12 54.62 2.25 2.12 2.37 21.8720.25 21.75 StD 7.02 9.86 9.59 0.70 0.83 0.51 2.85 3.28 3.80 Max 75 7066 3 3 3 25 26 28 Min 55 37 35 1 1 2 16 16 17 StE 2.48 3.48 3.39 0.250.29 0.18 1.00 1.16 1.34 Sum 506 449 437 18 17 19 175 162 174 McGill Day1: 87.37; Day 5: 78.49; Day 10: 78.84 Score

Less improvement was observed on the Affective aspects, and no effectwas observed on the Evaluative part of the Questionnaire. Following thistrend, treatment with 70 mg of RBE resulted in 41% reduction of pain asevaluated by Sensory part, but the total score of McGill Questionnairefor this experimental group yielded a 33% reduction after day 10. It isalso interesting to notice that 5 days of treatment with 70 mg of RBEalready resulted in 33% reduction of pain (total McGill score). Thisindicates that treatment with RBE may provide a moderately rapid effect(although not as acute as the effects of painkiller drugs such asNSAIDS). Exit interviews of study participants revealed that the firstsubjective improvements in pain were noticed after 3 days of thetreatment. This observation strongly suggests that a 3 day time-pointshould be included in any future RBE clinical efficacy study protocol tofollow rapid activity and effect on improvement of OA pain-relatedconditions.

Blood Chemistry Analysis.

Standard serum biochemistry analysis was performed on each serumcollected at day 1 and day 10. No unusual changes were noticed in anyparameters. All parameters were within normal range (data not shown).

Subjective Energy Tests.

In parallel to Mc-Gill Questionnaire, all participants were required toanswer 4 questions pertaining to their physical strength rate (Q1),mental alertness (Q2), physical endurance rate (Q3) and mood rate (Q4).Rates for all these questions were scaled 1-4. This Questionnaire(described in this article as Energy Score) was performed at day 1, 5and 10. The highest number indicated a generally elevated level of thefeeling rate.

All detailed data of this Questionnaire are presented in Table 6. Thesedata show that all participants reported feelings of increased mentalalertness, strength, endurance, and mood in the dose dependent mannerafter treatment with RBE. Treatment with 70 mg resulted in 122%improvement over Day 1 (Table 6), whereas the treatment with RBE at dose100 mg resulted in 81% improvement, indicating that treatment with thelower dose of 70 mg was optimal. Therefore, as it was noticed whenanalyzing McGill data, 70 mg dose seems to be the most potent forimprovement of parameters listed in Table 6. Also, the lowest dose of 35mg still provided significant Energy Scores increases of up to 74% after10 days of the treatment. In comparison, the McGill data for the samedose resulted in only 11% improvement. This may indicate that a primaryeffect of RBE is to modulate feelings of energy, mood, endurance andawareness since dose as low as 35 mg caused improvement of Energy scoreup to 74%. These results were rather unexpected since the Energy Scorewas followed only as an additional subjective parameter to supplementthe McGill pain score.

TABLE 6 Day 1 Day 5 Day 10 Q1 Q2 Q3 Q4 Q1 Q2 Q3 Q4 Q1 Q2 Q3 Q4 Group 1Average 1.14 1.28 1.28 1.42 1.42 2.14 1.71 2.48 2.14 2.28 2.57 2.57 StD0.37 0.48 0.48 0.53 0.53 0.37 0.48 0.78 0.69 0.95 0.97 0.97 Max 2 2 2 22 3 2 4 3 4 4 4 Min 1 1 1 1 1 2 1 2 1 1 1 2 StE 0.14 0.45 0.45 0.20 0.200.14 0.18 0.29 0.26 0.35 0.36 0.36 Sum 9 9 9 10 10 15 12 17 15 16 18 18Energy Score Average Sum +/− StD 9.25 +/− 0.5 13.5 +/− 3.1 16.75 +/− 1.581% increase over day 1 Group 2 Average 1.12 1.12 1.25 1.12 2.00 2.002.25 2.00 2.37 2.65 2.50 2.50 StD 0.35 0.35 0.46 0.35 1.30 1.06 0.880.00 1.40 1.18 0.92 0.92 Max 2 2 2 2 4 4 4 2 4 4 4 4 Min 1 1 1 1 1 1 1 21 1 2 2 StE 0.12 0.12 0.16 0.12 0.46 0.37 0.31 0.00 1.40 1.18 0.32 0.32Sum 8 9 10 9 16 16 18 16 19 21 20 20 Energy Score Average Sum +/− StD9.0 +/− 0.81 16.5 +/− 1.0 20.0 +/− 0.81 122% over Day 1 Group 3 Average1.14 1.28 1.28 1.57 1.57 1.42 1.71 2.14 2.00 2.14 2.42 2.14 StD 0.370.48 0.48 0.53 0.53 0.53 0.48 0.69 0.5 0.37 0.97 0.69 Max 2 2 2 2 2 2 23 3 3 4 3 Min 1 1 1 1 1 1 1 1 1 2 1 1 StE 0.14 0.18 0.18 0.18 0.20 0.200.18 0.26 0.21 0.14 0.36 0.26 Sum 8 9 9 11 11 10 12 15 14 15 17 15Energy Score Average Sum +/− StD 8.75 +/− 1.5 12.0 +/− 2.16 15.25 +/−1.25 77% over Day 1

Data collected on the Energy and McGill scores shows that RBE may indeedprovide relief for conditions associated with OA. According to theworking hypothesis mentioned at the beginning of this article, betalainsmay improve OA conditions due to their inhibitory effect on thechlorination of protein by hypochlorous acid released from activatedneutrophils. This hypothesis was based on two rationales: 1.) thatbetalains can reduce the amount of hypochlorous acid generated byactivated neutrophils; and 2.) that chlorinated proteins may contributeto onset of Osteoarthritis and associated conditions.

In order to begin testing this hypothesis, the serum levels of AdvancedOxidation Protein Products (AOPP) were measured using a commercial kit(Cell Biolabs, Inc., #STA318). This assay measures serum proteinsmodified by chloramine or hypochlorous acid. The detailed collected dataare summarized in Table 7. The data show significant broad range (Maxand Min) in baseline of AOPP at day 1 in each experimental group.Interestingly, this range was significantly reduced in each group after10 days of the treatment. Resulting Sum data show 36.3, 47.6 and 30.9%reduction in groups 1, 2 and 3, respectively. However, due to the broadrange of AOPP values at day 1, StD is relatively high. Consequently,these results are presented herein only as indicative and as ajustification for further clinical investigation on OA subjects withincreased serum AOPP levels.

TABLE 7 Group 1 Group 2 Group 3 Result Day 1 Day 10 Day 1 Day 10 Day 1Day 10 Average 20.9 13.2 21.4 12.8 19.2 15.21 Std 14.9 7.2 12.4 7.3 8.04.9 Max 50.8 23.7 44.0 25.4 33.9 25.4 Min 7.8 6.8 8.7 5.0 8.5 10.2 Sum144.6 92.2 171 89.7 154 106.5 % Reduction in Average values 36.9 50.220.8 % Reduction in Sum values 36.3 47.6 30.9

AOPP is known as a pro-inflammatory factor and inducer of TNF-alpharelease from monocytes. Therefore it was reasonable to verify whethertreatment with RBE may result in reduction of blood TNF-alpha levels. Inorder to further investigate possible actions of betalains, sera fromvolunteers treated with RBE were subjected to a Cytokines and Chemokinesarray as offered by Qynsys Inc. Collected data showed that prior totreatment only 10 participants out of 24 were found to have TNF-alphaabove the detection limit of 1 pg/mL per Elisa assay. However, treatmentwith RBE caused reduction of TNF-alpha in these 10 subjects after 10days of the treatment (Table 8). The same sera were additionally testedfor changes in the levels of other cytokines and chemokines. Thisscreening yielded data showing that treatment with RBE reduced serumlevel of IL6, GRO-alpha, and RANTES levels after 10 days of thetreatment. (Table 8). H

TABLE 8 3 5 1 2 TNF- 4 GRO- 6 Subject Group alpha IL-6 alpha RANTES 1 11.25 1.77 318.8 251442 1.00 1.62 235.0 218310 2 30.24 146.9 123.5 3754228.43 135.9 64.6 27109 3 2.76 3.9 257.5 36127 1.16 2.2 227.0 33248 DOD128.0% 23.7% 29.0% 16.6% N 3 3 3 3 4 2 3.05 ND 113.5 23800 1.42 26.710099 5 1.43 6.65 70.9 30582 1.37 5.70 51.3 23821 6 2.24 2.59 504.3380997 1.11 1.86 306.9 211381 7 116.07 476.2 120.2 27932 80.99 379.519.4 25647 DOD1 35.0% 22.0% 57.2% 33.7% N 4 3 4 4 8 3 1.34 4.7 73.315252 1.12 3.3 63.9 13115 9 1.70 2.2 48.5 36672 1.61 1.2 32.4 17600 10 172.35 293.2 135.2 15826 171.88 268.5 116.5 10265 DOD1 8.3% 28.3% 21.0%34.0% N 3 3 3 3

These data indicate that RBE may have favorable effect on blood levelsof TNF-alpha, IL-6, RANTES, and GRO-alpha. Due to rather limited numberof participants per group showing serum level of TNF-alpha higher than 1pg/mL, data in Table 8 are presented as indicative rather than definite.Here, the effect of RBE is shown on blood level of selected cytokinesand chemokines in subjects with TNF-alpha blood level >1 pg/mL. Allother cytokines and chemokines are presented at concentration pg/mL.Upper number in raw represents level of measured peptide at day 1,bottom number at day 10. DOD1=change in peptide level at day 10 over day1 and expressed as % of change

It should be noted that subjects with initial serum levels of TNF-alphabelow 1 pg/mL also reported reduction of McGill scores and improvementson Energy. This observation suggests that RBE may improve McGill andEnergy score in OA subjects in a TNF-alpha-independent manner.Measurement of blood levels of selected cytokines and chemokinesindicated that TNF-alpha, IL-6, GRO-alpha, and RANTES could be reducedafter 10 days of the treatment. It was also observed that initial bloodlevels of AOPP were reduced during 10 days of the treatment, confirmingthat betalains present in RBE may reduce detrimental effect ofhypochloric acid in human subjects.

Sinusitis, contact dermatitis, and acne tests were performed byadministration of the betalain compositions as reported for OA aboveusing daily dosage of 70 mg of the betalain composition. All subjectsreported subjective results on a bar scale and measurements were takento digitize the response as known in the art.

For treatment of sinusitis, Table 9 below illustrates exemplary resultsfor various listed parameters:

TABLE 9 Overall % Subject # and Score at Score at Improvement atInitials Question day 1 Day 7 day 7 #1: PTC Intensity of nasal drainage44 38 14 Age: 31 Intensity of headache 76 4 95 Gender: M Intensity offacial pain-pressure 4 2 50 Weight: 175 Intensity of watery/red eyes 12137 70 Height: 5′11 Intensity of itchy/palate nose 94 71 25 Case:Sinusitis Intensity of nasal congestion 95 44 54 Fatigue level 31 8 75Intensity of skin itching 71 31 57 Intensity of skin dryness 74 3 96Average improvement 59.6% #1: LS Intensity of nasal drainage 68 1 99Age: 24 Intensity of headache 33 1 97 Gender: F Intensity of facialpain-pressure 15 8 47 Weight: 208 Intensity of watery/red eyes 1 1 0Height: 5′7″ Intensity of itchy/palate nose 135 124 9 Case: SinusitisIntensity of nasal congestion 114 31 73 Fatigue level 131 66 50Intensity of skin itching 104 106 1 Intensity of skin dryness 135 135 0Average Improvement 41.7

For treatment of eczema and contact dermatitis, Table 10 belowillustrates exemplary results for various listed parameters:

TABLE 10 Treatment of subjects suffering Chronic skin allergy VisualAnalog Score Evaluation Overall % Subject # Score at Score at Score atImprovement at and Initials Question day 1 Day 7 Day 14 day 7 and 14 #1:GVR Intensity of skin redness in affected area 81 17 11 80 & 87 Age: 38Intensity of itching in the morning 4 4 4 0 Gender: M Intensity ofitching at bedtime 5 5 5 0 Weight: 215 Average size of red spots 26 11 858 & 70 Height: 6′2 Intensity of skin pain in affected areas 6 5 6 0Case: Intensity of skin dryness in affected areas 19 11 17 43 & 11Dermatitis Intensity of your nervousness 8 6 7 0 Frequency of allergicskin reactions 13 6 6 54 & 54 Intensity of skin tension in affected area13 6 5 54 & 62 Average Improvement after day 7 and 14 26.5 & 31.5 #1: NSIntensity of skin redness in affected area 28 10 NT 65 Age: 46 Intensityof itching in the morning 128 2 NT 98 Gender: F Intensity of itching atbedtime 128 2 NT 98 Weight: 183 Average size of red spots 40 2 NT 95Height: 5′8″ Intensity of skin pain in affected areas 4 2 NT 50 Case:Intensity of skin dryness in affected areas 12 4 NT 67 Eczema Intensityof your nervousness 96 65 NT 33 Frequency of allergic skin reactions 982 NT 98 Intensity of skin tension in affected area 14 2 NT 86 AverageImprovement 76.7

For treatment of acne, Table 11 below illustrates exemplary results forvarious listed parameters:

TABLE 11 Overall % Subject # and Score at Score at Improvement atInitials Question day 1 Day 7 Day 7 and 14 #1: BS Size of acne Pinhead-Pinhead, 50 Age: 11 lesions 2 mm 1 mm Gender: M # of lesions on 25 5 75Weight: 103 face Height: 5′4″ # of lesions on 0 0 0 neck #of lesions on27 12 56 chest # of lesions on 5 1 80 the back Average 52.2 improvement#2: SH Size of Acne 3-5 mm 2-3 mm 38 Age: 27 lesions Gender: F # oflesions on 4 2 50 Weight: 145 face Height: 5′1″ # of lesions on 4 2 50neck # of lesions on 0 0 0 chest # of lesions on 2 1 50 back Average37.6 Improvement

Thus, specific embodiments and applications of treatment of variousconditions using betalain-containing compositions have been disclosed.It should be apparent, however, to those skilled in the art that manymore modifications besides those already described are possible withoutdeparting from the inventive concepts herein. The inventive subjectmatter, therefore, is not to be restricted except in the spirit of theappended claims. Moreover, in interpreting both the specification andthe claims, all terms should be interpreted in the broadest possiblemanner consistent with the context. In particular, the terms “comprises”and “comprising” should be interpreted as referring to elements,components, or steps in a non-exclusive manner, indicating that thereferenced elements, components, or steps may be present, or utilized,or combined with other elements, components, or steps that are notexpressly referenced.

What is claimed is:
 1. A nutraceutical stimulant that enhances physicalstrength or physical endurance of an individual, comprising: adehydrated complex betalain mixture comprising a plurality of distinctbetalains from a starting material, wherein the distinct betalains havea near natural composition and are present in the mixture at a totalbetalain concentration of at least 4 wt %; wherein the dehydratedcomplex betalain mixture further comprises a plurality of sugars,wherein a ratio of the plurality of distinct betalains to the one ormore sugars is at least 0.3; a solid edible carrier; and wherein thestimulant is formulated for oral administration in a dosage unit thatprovides at least 70 mg of the mixture to thereby enhance the physicalstrength or physical endurance as measured by a subjective strength andendurance metric scale.
 2. The nutraceutical stimulant of claim 1,wherein the starting material is selected from a group consisting of abeet juice, a raw beet or a portion of the raw beet, a beet processingwaste liquid, a beet root culture or culture supernatants, a plantmaterial comprising one or more betalains, and a near natural betalainproduct.
 3. The nutraceutical stimulant of claim 1, wherein the totalbetalain concentration is between 4-20 wt %.
 4. The nutraceuticalstimulant of claim 1, wherein the dehydrated complex betalain mixturefurther comprises a plurality of sugars, wherein a ratio of theplurality of distinct betalains to the one or more sugars is at least0.3.
 5. The nutraceutical stimulant of claim 1, wherein the dosage unitprovides between 10 and 50 mg of total betalains.
 6. The nutraceuticalstimulant of claim 1, further comprising at least one of a non-steroidalanti-inflammatory drug and a cylooxygenase (COX) inhibitor.
 7. Adehydrated complex betalain mixture, comprising: a plurality of isolateddistinct betalains at a total betalain concentration of at least 4 wt %of the mixture; wherein the plurality of isolated distinct betalainshave a near natural composition with respect to a starting material fromwhich the betalains were obtained; wherein the dehydrated complexbetalain mixture further comprises a plurality of sugars at a ratio oftotal betalains to total sugars of at least 0.3; and wherein theisolated distinct betalains are formulated for oral administration in adosage unit that provides at least 70 mg of the mixture to therebyenhance the physical strength or physical endurance as measured by asubjective strength and endurance metric scale.
 8. The complex betalainmixture of claim 7, wherein the plurality of isolated distinct betalainscomprises at least ten distinct betalains.
 9. The complex betalainmixture of claim 7, wherein the starting material is selected from agroup consisting of a beet juice, a raw beet or a portion of the rawbeet, a beet processing waste liquid, a beet root culture or culturesupernatants, a plant material comprising one or more betalains, and anear natural betalain product.
 10. The complex betalain mixture of claim7, wherein the total betalain concentration is at least 20 wt %.
 11. Thecomplex betalain mixture of claim 7, wherein the ratio of the pluralityof distinct betalains to the total sugars is at least
 20. 12. Thecomplex betalain mixture of claim 7, having a residual water content ofequal or less than 8 wt %.
 13. The complex betalain mixture of claim 7,wherein the mixture is formulated in a dosage unit for oraladministration that provides at least 100 mg of the mixture to therebyenhance the physical strength or physical endurance as measured by asubjective strength and endurance metric scale.